Sodium influx in human leucocytes: a novel technique for assessment of sodium-proton antiport activity.

1. A triple-isotope method for measuring initial 22Na uptake rates in unactivated human leucocytes in the presence of serum, using 3H2O, [14C]sucrose and 22Na, is described. 2. This 22Na influx is inhibited by amiloride, with half-maximal inhibition at a concentration of 10(-5) mol/l. 3. About 70% of the 22Na influx in normal human leucocytes is inhibited by 1 mmol/l amiloride, with an external Na+ concentration of 10 mmol/l. 4. External Na+ antagonizes this inhibition by amiloride. The Km for external Na+ is 9 mmol/l. 5. Intracellular acidification from either external Na+ depletion or ammonium chloride incubation leads to an activation of amiloride-sensitive Na+ influx. 6. The amiloride-sensitive Na+-H+ antiport provides the major influx pathway for Na+ in unactivated human leucocytes in the presence of serum.